. In the mammalian testis, spermatogenesis takes place in the epithelium of the seminiferous tubule, the functional unit that produces millions of spermatozoa daily. The blood-testis barrier (BTB), located near the basement membrane (BM) of the seminiferous epithelium, confers gate-keeper and the fence function, which restricts the paracellular and transcellular transport of harmful substances such as toxicants across the barrier, respectively, to maintain spermatogenesis. The BTB also physically divides the seminiferous epithelium into a basal and an adluminal compartment;thus, meiosis I and II, and post-meiotic spermatid development take place in a specialized microenvironment (i.e., the adluminal compartment) behind the BTB. The BTB is one of the tightest blood-tissue barriers in mammals since cell junctions at the BTB are reinforced by bundles of actin filaments that lie perpendicular to apposing Sertoli cell plasma membranes, which are only found in the testis. However, the BTB is a dynamic structure, undergoing extensive restructuring at stage VIII of the epithelial cycle so that preleptotene spermatocytes can cross the BTB and enter the adluminal compartment for further development. It is now well established that a functional BTB is crucial for spermatogenesis. Studies completed in the past grant period have shown that BTB function is regulated by biologically active peptides released (i) at the apical ectoplasmic specialization (apical ES, a testis-specific adherens junction at the Sertoli-spermatid interface), and (ii) at the basement membrane (BM) mediated by a functional axis designated the apical ES-BTB-BM axis. In short, biologically active fragments of laminin and collagen chains released at the apical ES and the BM, respectively, act as autocrine factors to regulate BTB function in this axis. Herein, we will continue this important research to understand the underlying molecular mechanism(s) that regulates BTB function. First, we will unravel the mechanism(s) by which these fragments regulate Sertoli cell adhesion and BTB restructuring via their effects on the F-actin network at the BTB, such as actin binding and regulatory proteins. We will also examine the involvement of endosome-mediated protein trafficking and small regulatory RNAs in these events using state-of-the-art techniques and approaches. Second, we will identify the biological significance of biomolecules, such as cytokines, steroids and matrix metalloproteases, in regulating the stage-specific production of these bioactive peptides during the epithelial cycle of spermatogenesis. These findings will have a significant impact in understanding the biology of BTB in spermatogenesis. More important, these findings will shed new light on the development of novel reversible non-hormonal contraceptives for men. For instance, peptides derived from the apical ES and BM can be developed into contraceptives to perturb BTB function and impair spermatogenesis. Additionally, these findings can lead to the development of new therapeutic approaches to interfere with toxicant-induced male reproductive dysfunction, which is one of the leading causes of the decline in male fertility. For instance, the environmental toxicant cadmium mediates its effects at the BTB to induce male reproductive dysfunction;these findings provide crucial information in the design and synthesis of small molecules that either reverse or block cadmium-induced BTB disruption that leads to male infertility.

Public Health Relevance

Public Health Relevance Male infertility is on the rise in developed and developing countries across the world, which is partly attributed to environmental toxicant-induced male reproductive dysfunction. Studies have shown the blood-testis barrier (BTB) to be the primary target of these toxicants, such as cadmium, phthalates and bisphenol A. This project seeks to understand the molecular mechanism(s) of BTB regulation by biologically active peptides released at the apical ectoplasmic specialization (apical ES) and the basement membrane (BM) that regulate BTB dynamics in the apical ES-BTB-BM axis during the epithelial cycle of spermatogenesis. It also investigates the endogenous production of these regulatory peptides in staged tubules under the influence of cytokines, steroids and metalloproteases. These findings will provide a thorough understanding of the biology of the BTB and will provide new insight on the development of therapeutic approaches that protect the testis from harmful toxicants. Furthermore, this project will identify new targets for male contraceptive development. For instance, biologically active peptides can be developed to disrupt BTB function and impair spermatogenesis reversibly.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
2R01HD056034-06
Application #
8613215
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Moss, Stuart B
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Population Council
Department
Type
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10017
Chen, Haiqi; Mruk, Dolores D; Xia, Weiliang et al. (2016) Effective Delivery of Male Contraceptives Behind the Blood-Testis Barrier (BTB) - Lesson from Adjudin. Curr Med Chem 23:701-13
Tang, Elizabeth I; Mruk, Dolores D; Cheng, C Yan (2016) Regulation of microtubule (MT)-based cytoskeleton in the seminiferous epithelium during spermatogenesis. Semin Cell Dev Biol 59:35-45
Venkatesh, Deepak; Mruk, Dolores; Herter, Jan M et al. (2016) AKAP9, a Regulator of Microtubule Dynamics, Contributes to Blood-Testis Barrier Function. Am J Pathol 186:270-84
Li, Nan; Mruk, Dolores D; Lee, Will M et al. (2016) Is toxicant-induced Sertoli cell injury in vitro a useful model to study molecular mechanisms in spermatogenesis? Semin Cell Dev Biol 59:141-156
Gao, Ying; Lui, Wing-Yee; Lee, Will M et al. (2016) Polarity protein Crumbs homolog-3 (CRB3) regulates ectoplasmic specialization dynamics through its action on F-actin organization in Sertoli cells. Sci Rep 6:28589
Li, Nan; Mruk, Dolores D; Chen, Haiqi et al. (2016) Rescue of perfluorooctanesulfonate (PFOS)-mediated Sertoli cell injury by overexpression of gap junction protein connexin 43. Sci Rep 6:29667
Chen, Haiqi; Mruk, Dolores D; Lee, Will M et al. (2016) Planar Cell Polarity (PCP) Protein Vangl2 Regulates Ectoplasmic Specialization Dynamics via Its Effects on Actin Microfilaments in the Testes of Male Rats. Endocrinology 157:2140-59
Tang, Elizabeth I; Lee, Will M; Cheng, C Yan (2016) Coordination of Actin- and Microtubule-Based Cytoskeletons Supports Transport of Spermatids and Residual Bodies/Phagosomes During Spermatogenesis in the Rat Testis. Endocrinology 157:1644-59
Wen, Qing; Cheng, C Yan; Liu, Yi-Xun (2016) Development, function and fate of fetal Leydig cells. Semin Cell Dev Biol 59:89-98
Jesus, Tito T; Oliveira, Pedro F; Silva, Joaquina et al. (2016) Mammalian target of rapamycin controls glucose consumption and redox balance in human Sertoli cells. Fertil Steril 105:825-833.e3

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